[Leish-l] inquiry

Chang, Kwang-Poo KwangPoo.Chang at rosalindfranklin.edu
Wed Jun 15 06:24:37 BRT 2011


I did a search for L donovani LV9 and found its origin from Ethiopia (MHOM/ET/67/HU3). 
 
I wonder about its genetic or genomic similarity to L donovani Sudan strains and Indian L donovani. Correct me if I should be wrong about my suspicion that these three don't give you a distinct clade or subclade within the L donovani/infantum complex. This is relevant to the definition of your "L donovani/L infantum hybrid".
 
KP
 
 

________________________________

From: leish-l-bounces at lineu.icb.usp.br on behalf of Petr Volf
Sent: Thu 6/9/2011 5:55 AM
To: BobKillick-Kendrick; Patrick Bastien
Cc: leish-l at lineu.icb.usp.br; Lachaud; Ravel; Pratlong F
Subject: Re: [Leish-l] inquiry


Dear Bob, 
 
hybridization takes place in the vector but I am not sure if our finding in Cukurova brings a supporting evidence for this. I can imagine also other scenarios, for example there are seasonal workers in this area, man comming back from army servis etc. 
 
Experimentally we confirmed that at least two local vectors, P. perniciosus and P. tobbi, are highly susceptible to both, "typical" L. donovani (LV9) as well MON1 L. infantum. They are, of course, highly susceptible to the CUK strains and transmit them into mice (Carla Maia presented this in ISOPS this April). For me this is not surprising as P. perniciosus and P. tobbi are Larroussius species and permissive vectors.
 
Best wishes
Petr
PS: Just to answer briefly older question about hybrids (I sent this reply before only to Patrick as I feel that discussion is becomming spread too much): based on techniques described in the manuscript we originally thought that these strains are L. infantum (there is nothing about hybrids in the paper). Recently we obtained new data including the whole genome sequence and this allow me to mention that the CUK strain is probably a hybrid.



	----- Original Message ----- 
	From: BobKillick-Kendrick <mailto:killickendrick at wanadoo.fr>  
	To: Patrick Bastien <mailto:patrick.bastien at univ-montp1.fr>  ; Petr Volf <mailto:volf at cesnet.cz>  
	Cc: Pratlong F <mailto:f-pratlong at chu-montpellier.fr>  ; Lachaud <mailto:laurence.lachaud at chu-nimes.fr>  ; Ravel <mailto:christophe.ravel at univ-montp1.fr>  ; leish-l at lineu.icb.usp.br 
	Sent: Wednesday, June 08, 2011 5:59 PM
	Subject: Re: [Leish-l] inquiry

	If there are hybrids between infantum and donovani in Turkey, the cross must have taken place in a vector. Petr, is there a sand fly species there that transmits both parasites? If not, how can there be a hybrid?
	    Bob

		----- Original Message ----- 
		From: Patrick Bastien <mailto:patrick.bastien at univ-montp1.fr>  
		To: Petr Volf <mailto:volf at cesnet.cz>  
		Cc: Pratlong F <mailto:f-pratlong at chu-montpellier.fr>  ; Lachaud <mailto:laurence.lachaud at chu-nimes.fr>  ; Ravel <mailto:christophe.ravel at univ-montp1.fr>  ; leish-l at lineu.icb.usp.br 
		Sent: Friday, June 03, 2011 3:37 PM
		Subject: Re: [Leish-l] inquiry

		Dear Petr,
		
		Two comments :
		1) I am curious to know how you define a hybrid between L. infantum  
		and L. donovani, knowing that these species are quite difficult to  
		differentiate by simple sequence analysis. Is it using the MLST method  
		described in your paper attached ? Are the genes you are using single  
		copy or duplicated ? Have you used isoenzymes for confirmation ?
		
		2) Talking of such, I am always puzzled when I see efforts for  
		correlating molecular and isoenzyme-based phenogramms (Fig. 4, your  
		article attached). Worse, your text clearly states that the strains  
		analyzed are "unambiguously distinct from the MON-1 zymodeme (...) of  
		L. infantum". This is particularly risky, as it is to use zymodeme  
		names (MON-...) in a phylogenetic reconstruction using other intrinsic  
		(here molecular) criteria. Indeed, a zymodeme may include different  
		genotypes and a genotype may theoretically cover several zymodemes. A  
		zymodeme is nothing else than an "operational taxonomic unit" used for  
		classification (phenetics) (see the famous paper by Rioux et al.  
		attached); as such, it has no biological nor evolutionary signification.
		Similarly, when you write "Among accessible isolates typed using the  
		MLST method, our strain is close to MON-188, the L. infantum strain  
		isolated by Gramiccia (Pratlong et al., 2003)(...)", the MON-... is  
		abusively assimilated to a given strain (here ISS800), whereas one  
		zymodeme normally clusters several strains, that might be found  
		clustered differently using your method or another method.
		Another important point is that the MON-... is defined by the  
		examination of a series of 15 given iso-enzymes, but this is also  
		dependent upon the technique used (and, to a lesser degree, to the  
		person/research group who defines it). That is why Rioux et al.  
		recommended that each identifying center use its own code (such as  
		MON- for Montpellier or LON- for London). Indeed, MON-188 in  
		Montpellier migh tbe different from MON-188 in another center if the  
		system used is not exactly the same.
		All this may seem trivial, but there is a real danger, as it slowly  
		introduces in the minds of our colleagues scientists that different  
		classifications are equivalent.
		I know that I am touching on a question which can yield hours of  
		debate and thousands of e-mails; so, although I am sure you will want  
		to reply to this, please note that I do not want to lauch such a  
		debate online !
		
		Best wishes
		Patrick
		
		Professeur Patrick Bastien
		Laboratoire de Parasitologie-Mycologie, Faculté de Médecine
		UMR MIVEGEC (CNRS 5290 - IRD 224 - Université Montpellier 1)
		Centre National de Référence des Leishmania
		CHU de Montpellier
		39 Avenue Charles Flahault
		34295 Montpellier Cedex 5, France
		
		Petr Volf <volf at cesnet.cz> a écrit :
		
		> Re: [Leish-l] inquiryHi K.P.,
		> in Cukurova region, Turkey, cutaneous L. infantum (now it seems that  
		>  we are dealing with L.donovani/L. infantum hybrid) grew very  
		> poorely  if we isolated them from patients. Only 1 of 25 isolations  
		> was  succesfull. However, the same strain (confirmed by molecular   
		> methods) grew repeatedly and very well if we isolated them from   
		> sandflies. It might be useful for Dr. Sharma try to get isolates   
		> from sand flies. It is very laborious but very useful, in Cukurova   
		> we got about dozen of isolates by this method (all identical).
		> Patients are rK39 negative, see attached paper.
		> Best wishes
		> Petr
		>   ----- Original Message -----
		>   From: Chang, Kwang-Poo
		>   To: Sharman ; Hiro Goto ; elfadil abass
		>   Cc: leish-l at lineu.icb.usp.br
		>   Sent: Sunday, May 22, 2011 1:39 AM
		>   Subject: Re: [Leish-l] inquiry
		>
		>
		>   Dr. NL Sharma has been working on an important CL endemic area   
		> along the Satluj River valley to the south of Himalaya in India   
		> (Please correct me should I be wrong for anything I said here and   
		> below). I had the good fortune of visiting the site several years   
		> back courtesy of Dr. Sharma.
		>
		>   I believe Dr. Sharma's finding is important, since the parasites   
		> there are different from the familiar Indian L donovani in Bihar.   
		> The parasites are refratory to in vitro cultivation. They do   
		> differentiate into promastigotes and may grow a little, but  can't   
		> really be subcultured to establish passageable lines. This is very   
		> much reminescent of L infantum in the Mediterranean area. I recall   
		> Dr. Sharma has also found rK39+ dogs, if I remember correctly. If   
		> so,  Satluj river valley is endemic to the infantile CL.
		>
		>   Analyses of several batches of basically clinical CL samples from   
		> Dr. Sharma showed evidence of L infantum, but also L tropica as well  
		>  as a mixture of the two in one sample. This is the same picture we   
		> have noted for samples from Hatay, Turkey.
		>
		>   These observations make me wonder a lot about our current   
		> knowledge on the clinico-epidemiology based on data collected   
		> previously by analyses of cultured promastigotes from one or few   
		> 'representative samples'. Nowaday, technology makes it very doable   
		> to work with biological samples for Leish DNAs directly from sand   
		> flies, patients and reservoir animals.
		>
		>   KP
		>
		>
		>
		> ------------------------------------------------------------------------------
		>   From: leish-l-bounces at lineu.icb.usp.br on behalf of Sharman
		>   Sent: Fri 5/20/2011 11:31 PM
		>   To: Hiro Goto; elfadil abass
		>   Cc: leish-l at lineu.icb.usp.br
		>   Subject: Re: [Leish-l] inquiry
		>
		>
		>   Dear all
		>   I agree with Hiro gito, We are working on a focus where the CL is
		>   predominantly caused by L. donovani, and the rK 39 STRIPS GIVE POSITIVE
		>   RESULTS WITH SERA as well as serous exudate from the lesion. The   
		> results are
		>   dependent upon species.
		>   NL Sharma
		>
		>   --------------------------------------------------
		>   From: "Hiro Goto" <hgoto at usp.br>
		>   Sent: Wednesday, May 18, 2011 12:09 AM
		>   To: "elfadil abass" <elfadil_abass at yahoo.com>
		>   Cc: <leish-l at lineu.icb.usp.br>
		>   Subject: Re: [Leish-l] inquiry
		>
		>   > Dear all,
		>   > In our oppinion, DAT and rK39 for those samples are not indicated  since
		>   > these tests are produced for the diagnosis of visceral  leishmaniasis.
		>   > In case of tegumentary leishmaniasis, it is very appropriate the
		>   > observation of J.J. Shaw appointing species specificity of antibody
		>   > response in these cases. Titers of antibodies are in general low  
		>  in  these
		>   > cases therefore depending on the species, it may result    
		> negative. We have
		>   > published a review recently in Expert Rev. Anti  Infect. Ther. 8(4),
		>   > 419?433 (2010), Current diagnosis and treatment of  cutaneous and
		>   > mucocutaneous leishmaniasis, where we raise this point.
		>   > Hiro Goto
		>   >
		>   > Citando elfadil abass <elfadil_abass at yahoo.com>:
		>   >
		>   >> Dear all I would recommend using DAT and rK39 strip test to measure
		>   >> antibody
		>   >> responses and to evaluate the diagnostic efficiency for both tests  in
		>   >> such group
		>   >> of patients.
		>   >>
		>   >>  Elfadil Abass
		>   >>   
		> ________________________________________________________________________________
		>   >> Institute of Medical Microbiology and Hospital Hygiene
		>   >> Philipps University Marburg
		>   >> BMFZ / Hans-Meerwein Straße 2
		>   >> D-35033 Marburg, Germany
		>   >>   
		> ________________________________________________________________________________
		>   >>
		>   >>
		>   >>
		>   >>
		>   >> ________________________________
		>   >> From: Nuha Nuwayri-Salti <racha at aub.edu.lb>
		>   >> To: saad saad <saad1426 at gmail.com>; "leish-l at lineu.icb.usp.br"
		>   >> <leish-l at lineu.icb.usp.br>
		>   >> Sent: Fri, May 13, 2011 9:39:33 AM
		>   >> Subject: Re: [Leish-l] inquiry
		>   >>
		>   >> Dear Saad first precaution to take is to separate your samples into
		>   >> several
		>   >> portions each (at least 5 each being no more than a few hundred(200-400)
		>   >> microliters). This is a necessary precaution to avoid freezing and
		>   >> thawing
		>   >> several times the same sample which will be the case should you do
		>   >> different
		>   >> studies at different times which is unavoidable.
		>   >>
		>   >> What you can do is correlate the type(ulcerated, abscess, furuncle
		>   >>  etc.. ) the
		>   >> locale, the number and age of lesions with the levels of   
		> antibody in  the
		>   >> sera of
		>   >> these patgients and also monitor cell mediated immunity with leishmanin
		>   >> skin
		>   >> test!
		>   >> I have just published (in print) an article about having circulating
		>   >> parasites
		>   >> in some of these patients with apparently pure cutaneous disease. It is
		>   >> the
		>   >> first paper that revealed this fact. It would be interesting to confirm
		>   >> or
		>   >> de-confirm this fact repeating what we did.
		>   >> Best wishes
		>   >>
		>   >> Nuha Nuwayri-Salti MD
		>   >>
		>   >>
		>   >>
		>   >> AOA
		>   >> Medical Honor Society
		>   >>
		>   >>
		>   >>
		>   >>
		>   >> -----Original Message-----
		>   >> From: leish-l-bounces at lineu.icb.usp.br
		>   >> [mailto:leish-l-bounces at lineu.icb.usp.br]
		>   >> On Behalf Of saad saad
		>   >> Sent: Friday, April 29, 2011 11:23 PM
		>   >> To: leish-l at lineu.icb.usp.br
		>   >> Subject: [Leish-l] inquiry
		>   >>
		>   >> Dear all
		>   >> Hi. i am Saad from Saudi Arabia. i have 60 sera from positive case of
		>   >> CL from south west of the country. I would like to have your
		>   >> recommendations to start a good research line in CL using these sera.
		>   >> Thanks in advance for your help
		>   >> _______________________________________________
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		>   >> Leish-l at lineu.icb.usp.br
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		>   >> Leish-l mailing list
		>   >> Leish-l at lineu.icb.usp.br
		>   >> http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l
		>   >>
		>   >
		>   >
		>   >
		>   > Profa. Dra. Hiro Goto
		>   > Laboratório de Soroepidemiologia e Imunobiologia
		>   > Instituto de Medicina Tropical de São Paulo, USP
		>   > Av. Dr. Enéas de Carvalho Aguiar, 470, prédio II, quarto andar
		>   > 05403-000 - São Paulo, SP
		>   > Tel. +55-11-3061 7023, 3061 7056 ou 3061 7027
		>   > Fax. +55-11-3061-8270
		>   >
		>   > _______________________________________________
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		>   >
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		>
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